HCC cells seeded on coverslips were washed 3 times with PBS, fixed with 4% paraformaldehyde for 15 min, and permeabilized with 0.3% Triton X-100 at room temperature for 40 min for nuclear proteins. Then, the cells were blocked with 5% BSA for 30 min and stained with primary antibodies (Additional file 1 : Table S2) overnight at 4 °C (Additional file 1 : Table S1). After washing with PBS and incubation with Alexa fluor 594-conjugated goat-anti rabbit secondary antibody (Proteintech) at dark room for 1 h, the cells were incubated with 0.1% 4′,6-diamidino-2-phenylindole (DAPI) for 5 min, washed with PBS, and then observed under an inverted fluorescence confocal microscope (Olympus).
Alexa fluor 594 conjugated goat anti rabbit secondary antibody
Manufactured by Proteintech
Sourced in United States, China
Alexa Fluor 594-conjugated goat-anti rabbit secondary antibody is a fluorescently labeled secondary antibody designed for use in various immunodetection techniques. It binds to rabbit primary antibodies, allowing for the visualization and detection of target proteins or antigens in samples.
Automatically generated - may contain errors
Lab products found in correlation
Inverted fluorescence confocal microscope, by Olympus (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
Inverted fluorescence microscope, by Olympus (1 mentions)
Coolsnap pro cf, by Media Cybernetics (1 mentions)
Glp 1r, by Santa Cruz Biotechnology (1 mentions)
Pparγ, by Santa Cruz Biotechnology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Citrate antigen retrieval solution, by Beyotime (1 mentions)
4 protocols using alexa fluor 594 conjugated goat anti rabbit secondary antibody
1
Immunofluorescence Staining of HCC Cells
2
Immunofluorescence Staining of HCC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HCC cells seeded on coverslips were 4% paraformaldehyde‐fixed for 20 min, permeabilized with 0.3% Triton X‐100 at room temperature (40 min for nuclear proteins, 20 min for cytosolic proteins or without this procedure for membrane proteins). Then, slides were blocked with 3% BSA for 1 h at room temperature, followed by incubation with primary antibodies (Table S2 ) overnight at 4°C. The next day, all the followed procedures were performed in the dark at room temperature. The slides were incubated with Alexa fluor 594‐conjugated goat‐anti‐rabbit secondary antibody (Proteintech) for 1 h, and the nucleus was stained with DAPI (Solarbio) for 10 min. Images were captured using an inverted fluorescence microscope (Olympus). The fluorescence intensity was evaluated using ImageJ software (NIH, USA).
3
TNFR2 Immunofluorescent Staining in MC3T3-E1 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunofluorescent staining, MC3T3-E1 cells were washed with cold PBS, fixed with 4% paraformaldehyde, permeabilized with 0.2% Triton X-100 in 1% bovine serum albumin for 30 min, blocked with 5% bovine serum albumin for 1 h at room temperature, and incubated overnight at 4 °C with polyclonal rabbit anti-TNFR2 antibody (1:100 dilution, 19,272–1-AP, Proteintech, USA). Cells were then incubated with Alexa Fluor 594-conjugated goat anti-rabbit secondary antibody (1:100 dilution, SA00006–4, Proteintech, USA). The cell nuclei were stained with 4, 6-diamino-2-phenylindole (DAPI). Stained MC3T3-E1 cells were visualized under a fluorescent microscope (Olympus, BX51, Japan), and images were captured by a CCD camera (CoolSNAP-Pro cf., Media Cybernetics, USA). The fluorescence intensity was counted in selected merged microscopic images by Image-Pro Plus 6.0 software.
4
Immunofluorescence Analysis of GLP-1R and PPAR-γ in Rat Brain
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh brains of rats were isolated and embedded by OTC (Sakura Finetek Japan Co., Ltd, Tokyo, Japan), and then, they were cut to get frozen 10μm sections. After slices were repaired in the high-pressure cooker by citrate antigen retrieval solution (Beyotime, Shanghhai, China, Catalogue No.: P0081) and blocked by 5% normal goat serum (Beyotime, Shanghai, China, Catalogue No.: C0265) (1 hour at room temperature). They were incubated with rabbit-anti-rat primary antibody to GLP-1R (Santa Cruz Biotechnology, Inc., Dallas, TX, USA, Catalogue No.: sc-390774) and PPAR-γ (Santa Cruz Biotechnology, Inc., Dallas, TX, USA, Catalogue No.: sc-390740) at 4° C overnight. After washing for 3 times, Alexa Fluor 594 conjugated goat-anti-rabbit secondary antibody (Proteintech, Wuhan, China, Catalogue No.: SA00013-3) was used to detect the primary antibody. Finally, sections were stained by DAPI (Beyotime, Shanghhai, China, Catalogue No.: P0131) and washed for 3 times with PBS. GLP-1R (PPAR-γ) and DAPI (in hippocampal dentate gyrus and cortex) were observed by a fluorescence microscope (Olympus, Japan) after triggered at 594 nm and 358 nm respectively. Image were captured by (CellSens Standard).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!